Method for the Production of Proteins
AssigneeNovo Nordisk HealthCare A/G
InventorsJanus Krarup, Egon Persson
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Abstract
The present invention relates to a process for the purification of a protease.
Claims48
A method for production of a purified GLA-residue containing serine protease, said method comprising:
- (i) culturing a host cell expressing said GLA-residue containing serine protease in a culture medium under conditions appropriate for expression of said GLA-residue containing serine protease;
- (ii) recovering all or part of the culture medium comprising said GLA-residue containing serine protease;
- and
- (iii) purifying said GLA-residue containing serine protease from the culture medium;
- wherein the pH under step (iii) has a value between 4.5 and 6.9 or has a value between 8.6 and 10 and the molar ratio of calcium ions and GLA-residue containing serine protease (Ca2+:
- GLA-residue containing serine protease) under step (iii) is lower than 0.50.
A process for the purification of a GLA-residue containing serine protease whereby a solution of a GLA-residue containing serine protease is subjected to a number of purification steps, wherein the molar ratio of calcium ions and GLA-residue containing serine protease (Ca2+:
- GLA-residue containing serine protease) is, at least in one of the purification steps, lower than 0.50.
A method for stabilizing a Factor VII polypeptide in a solution comprising the Factor VII polypeptide, whereby said solution comprising the Factor VII polypeptide is subjected to the steps of adding calcium to obtain a molar ratio of calcium ions and Factor VII polypeptide (Ca2+:FVII polypeptide) lower than 0.5 and adjusting the pH of the solution comprising a GLA-residue containing serine protease to a value between 4.5 and 6.9 or to a value between 8.6 and 10.
Dependencies are inferred from the claim text.